Contact Us Add to Favorite
 
NianQi Search Adv Search
33
农业生物技术学报  2019, Vol. 27 Issue (5): 856-863    DOI: 10.3969/j.issn.1674-7968.2019.05.009
Articles and Letters Current Issue | Archive | Adv Search |
Over-expression of Bovine (Bos taurus) BTN1A1 Gene Promotes Lipid Droplet Synthesis in Mammary Epithelial Cells
ZHANG Meng-Lu*, FAN Xiao-Na*, XING Zhi-Yang, WANG Yue-Ying, WANG Jiang, CHU Bei-Bei, HAN Li-Qiang**, YANG Guo-Yu**
College of Animal Science and Veterinary Medicine/Key Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture, Henan Agricultural University, Zhengzhou 450002, China
Download: PDF (3536 KB)   HTML (1 KB) 
Export: BibTeX | EndNote (RIS)      
Abstract  As a lipid droplet protein, butyrophilin subfamily 1 member A1 (BTN1A1) is associated with the formation and secretion of milk fat globules.In order to investigate the effect of over-expression bovine (Bos taurus) BTN1A1 on lipid droplets in mammary epithelial cells, the coding region of BTN1A1 gene was amplified by PCR, and Lenti-BTN1A1 lentivirus recombinant vector was constructed.The virus was packaged into recombinant vector and infected bovine mammary epithelial cells.BTN1A1 over-expression cell lines were selected by applying puromycin in cell culture medium.DNA was extracted from the over-expressed cell lines and genotypes were identified.Expression of BTN1A1 gene and protein was detected by qRT-PCR and Western blot.The effects of BTN1A1 over-expression on cell lipid droplets were analyzed by fluorescent labeling with Nile red.BTN1A1-pEGFP-N1 vector was constructed to observe the co-localization of BTN1A1 protein and lipid droplet.The results showed that the CDS sequence of the cloned BTN1A1 gene was about 1 580 bp.The lentiviral vector recombinant with BTN1A1 was constructed.BTN1A1 cell line was selecteded by 5 μg/mL purinomycin.Genotypic identification showed that exogenous BTN1A1 gene was successfully inserted into the cell genome.Compared with control group, the expression level of BTN1A1 gene mRNA was 489 times greater (P<0.001) ,while protein expression was also increased in over-expression BTN1A1 cell line.Fluorescence values of lipid droplets showed that over-expression BTN1A1 significantly increased lipid droplet content with the extension of cell culture time (P<0.01).BTN1A1 protein was found to co-localized with lipid droplets in cells.In this study, over-expression BTN1A1 promoted lipid droplet synthesis in mammary epithelial cells.This study provides basic data to elucidate the mechanism of lipid droplet synthesis in mammary epithalial cells of dairy cows.
Key wordsBTN1A1      Mammary epithelail cell      Over-expression      Lipid droplet     
Received: 20 August 2018     
ZTFLH:  S852.2  
Corresponding Authors: qlhan2001@126.com;haubiochem@163.com   
Service
E-mail this article
Add to my bookshelf
Add to citation manager
E-mail Alert
RSS
Articles by authors
ZHANG Meng-Lu
FAN Xiao-Na
XING Zhi-Yang
WANG Yue-Ying
WANG Jiang
CHU Bei-Bei
HAN Li-Qiang
YANG Guo-Yu
Cite this article:   
ZHANG Meng-Lu,FAN Xiao-Na,XING Zhi-Yang, et al. Over-expression of Bovine (Bos taurus) BTN1A1 Gene Promotes Lipid Droplet Synthesis in Mammary Epithelial Cells[J]. 农业生物技术学报, 2019, 27(5): 856-863.
URL:  
http://journal05.magtech.org.cn/Jwk_ny/EN/10.3969/j.issn.1674-7968.2019.05.009     OR     http://journal05.magtech.org.cn/Jwk_ny/EN/Y2019/V27/I5/856
Copyright © Editorial Board of 农业生物技术学报
Supported by:Beijing Magtech